Antibiotic resistance pattern and frequency of ESBL producing Enterobacteriaceae isolated from lettuce and spinach in Gorgan

مجله پژوهشهای علوم و صنایع غذایی ایران

Article 3, Volume 14, Issue 4 - Serial Number 52, September and October 2018, Pages 495-502   

Document Type: Research Article

DOI: 10.22067/ifstrj.v1396i0.62853

Authors

Anegol Kheradmand¹; Ania Ahani Azari^{* 1}; Ahmad Danesh²

¹Department of Microbiology, Gorgan Branch, Islamic Azad University, Gorgan, Iran.

²Department of Health and Social Medicine, School of Medicine, Golestan University of Medical Sciences, Iran.

Abstract

Introduction: In recent years, there have been many epidemic outbreaks caused by consumption of contaminated vegetables. In most cases, Enterobacteriaceae were responsible for these epidemics. The purpose of this study was to investigate antibiotic resistance pattern and frequency of strains producing broad-spectrum beta-lactamase (ESBL) isolated from lettuce and spinach produced in Gorgan, Iran.

Material and Methods: After culturing the isolates on MacConkey medium, colonies with specific morphological characteristics were selected. After preparation of pure culture, strains resistant to cefotaxime were identified and strains belonging to the Enterobacteriaceae family were isolated. The isolates were then studied for broad-spectrum beta-lactamases (ESBL) and their antibiotic resistance patterns were determined.

Results & Discussion: The results of this study showed that ESBL Enterobacteriaceae is present in vegetables such as lettuce and spinach. It was also found that all isolated ESBL Enterobacteriaceae from spinach samples were resistant to Cotrimoxazole, Nalidixic Acid, Tetracycline, Chloramphenicol, and Amoxicillin, and sensitive to Imipenem. Isolated ESBL Enterobacteriaceae from lettuce samples was sensitive to Amikacin and Imipenem. Multiple antibiotic resistance was observed in all isolates. Therefore, it can be concluded that use of animal fertilizers increases the risk of antibiotic-resistant bacteria in vegetables and ultimately in humanand animals.

Keywords

Enterobacteriaceae; broad-spectrum beta-lactamases; Vegetables; antibiotic resistance

References

Ali Yahya Saeed, Hayfaa Mazin, AwazArshad Saadi, Solin Omar Hussein (2013). Detection of Escherichia coli O157 in vegetables. IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) e-ISSN: 2319-2380, p-ISSN: 2319-2372.Volume 6, Issue 2 (Nov. – Dec). PP 16-18.

Bezanson, G.S., MacInnis, R., Potter, G. and Hughes, T (2008). Presence and potential for horizontal transfer of antibiotic resistance in oxidase-positive bacteria populating raw salad vegetables. International Journal of Food Microbiology vol. 127, no. 1-2, pp. 37–42.

Buchholz, U., Bernard, H., Werber, D., Böhmer, MM., Remschmidt, C., Wilking, H., Delere, Y., an der Heiden, M., Adlhoch, C., Dreesman, J., Ehlers, J., Ethelberg, S., Faber, M., Frank, C., Fricke, G., Greiner, M., Höhle, M., Ivarsson, S., Jark, U., Kirchner, M., Koch, J., Krause, G., Luber, P., Rosner, B., Stark, K., Kühne, M (2011). German outbreak of Escherichia coli O104:H4 associated with sprouts. New England Journal of Medicine 365(19):1763–1770.

Falomir, M.P., Gozalbo, D. and Rico, H (2010). Coliform bacteria in fresh vegetables: from cultivated lands to consumers. Current Research, Technology and education topics In Applied Microbiology and Microbial Biotechnology 1175-1181.

Jensen, AN., Storm, C., Forslund, A., Baggesen, DL., Dalsgaard, A (2013). Escherichia coli Contamination of Lettuce Grown in Soils Amended with Animal Slurry. Journal of Food Protection Vol. 76, No. 7, Pages 1137–1144.

Johan Tham, Mats Walder, Eva Melander, and Inga Odenholt (2012). Prevalence of extended-spectrum beta-lactamase-producing bacteria in food. Infection and Drug Resistance 5 143–147.

Kevin Holvoet, Imca Sampers, Benedicte Callens, Jeroen Dewulf, Mieke Uyttendaele (2013). Moderate prevalence of antimicrobial resistance in Escherichia coli isolates from lettuce, irrigation water, and soil. Journal of Applied & Environmental Microbiology 79: 6677–6683.

Magiorakos, AP., Srinivasan, A., Carey, RB., Carmeli, Y., Falagas, ME., Giske, CG., Harbarth, S., Hindler, JF., Kahlmeter, G., Olsson-Liljequist, B., Paterson, DL., Rice, LB., Stelling J, Struelens, MJ., Vatopoulos, A., Weber, JT., Monnet, DL (2012).Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clinical Microbiology and Infection Mar; 18(3):268-81.

Mohammad Khiyami, Noura AL-Faris, Basel Busaeed and Hassan Sher (2011). Food borne pathogen contamination in minimally processed vegetable salads in Riyadh, Saudi Arabia .Journal of Medicinal Plants Research 4 February, Vol. 5(3), pp. 444-451.

Mohammed Uddin Rasheed, Nooruddin Thajuddin, Parveez Ahamed P, Zelalem Teklemariam & Kaiser Jamil (2014). Antimicrobial drug resistance in strains of Escherichia coli isolated from food sources. Revista do Instituto de Medicina Tropical de São Paulo. Sao Paulo 56 (4):341-346.

Natasha Bhutani, Chithra Muraleedharan, Deepa Talreja,, Sonia Walia Rana, Sandeep Walia, Ashok Kumar, and Satish K.Walia1 (2015). Occurrence of Multidrug Resistant Extended Spectrum Beta-Lactamase-Producing Bacteria on Iceberg Lettuce Retailed for Human Consumption BioMed Research International Volume 2015, Article ID 547547, 10 pages.

Patrick M. K. Njage and Elna M. Buys (2014). Pathogenic and commensal Escherichia coli from irrigation water show potential in transmission of extended spectrum and AmpC β-lactamases determinants to isolates from lettuce. Microbial Biotechnology 462-473.

Puspanadan, S., Afsah-Hejri, L., John, Y. H. T., Rukayadi, Y., Loo, Y. Y., Nillian, E., Kuan, C. H., Goh, S. G., W. S. Chang, Lye, Y. L., Mohd Shahril, N., N. Yoshitsugu, M. Nishibuchi, Son, R ( 2013).Characterization of extended-spectrum β -lactamases (ESBLs) producers in Klebsiella pneumoniae by genotypic and phenotypic method. International Food Research Journal 20(3): 1479-1483.

Raphael, E., Wong, LK. Riley, LW (2011). Extended-spectrum betalactamase gene sequences in gram-negative saprophytes on retail organic and nonorganic spinach. Journal of Applied & Environmental Microbiology 77(5):1601–1607.

Raul Jesu´ s Mesa, Vanessa Blanc, Anicet R. Blanch, Pilar Corte´s, Juan Jose´ Gonza´ lez, Susana Lavilla, Elisenda Miro´, Maite Muniesa, Montserrat Saco, MaTeresa To´ rtola, Beatriz Mirelis, Pere Coll, Montserrat Llagostera, Guillem Prats and Ferran Navarro (2006). Extended-Spectrum-blactamse-producing Enterobacteriaceae in different environments (human, food, animal farms and sewage). Journal of Antimicrobial Chemotherapy 58:211–215.

Reuland, E.A., Naiemi, N. al., Raadsen, S. A., Savelkoul, P.H.M., Kluytmans, J.A.J.W., Vandenbroucke-Grauls, C.M.J.E (2014). Prevalence of ESBL-producing Enterobacteriaceae in raw vegetables. European Journal of Clinical Microbiology & Infectious Diseases 33:1843–1846.

Ruimy, R., Brisabois, A., Bernede, C., Skurnik, D., Barnat, S., Arlet, G., Momcilovic, S., Elbaz, S., Moury, F., Vibet, MA., Courvalin, P., Guillemot, D., Andremont, A (2010). Organic and conventional fruits and vegetables contain equivalent counts of Gram-negative bacteria expressing resistance to antibacterial agents. Environmental Microbiology 12(3):608– 615.

Sabry A. Hassan, Abdullah D. Altalhi, Youssuf A. Gherbawy, and Bahig A. El-Deeb (2011). “Bacterial load of fresh vegetables and their resistance to the currently used antibiotics in Saudi Arabia”. Foodborne Pathogens and Disease Volume 8, Number 9, 1011-1018.

Somayeh Mazaheri, Siavosh Salmanzadeh-Ahrabi, Tahereh Falsafi, Mohammad-Mehdi Aslani (2014). Isolation of Enteropathogenic Escherichia coli from lettuce samples in Tehran. Gastroenterology and Hepatology from Bed to Bench 7(1):38-42.

Sybille Boehme, Guido Werner, Ingo Klare, Rolf Reissbrodt and Wolfgang Witte (2004).Occurrence of antibiotic resistant enterobacteria in agricultural foodstuffs. Molecular Nutrition & Food Research 48:522–531.